GSK-3α/β and MEK inhibitors assist the microenvironment of tumor initiation

Ghmkin Hassan; Said M Afify; Maram H Zahra; Hend M Nawara; Kazuki Kumon; Yoshiaki Iwasaki; David S Salomon; Akimasa Seno; Masaharu Seno

doi:10.1007/s10616-023-00575-1

GSK-3α/β and MEK inhibitors assist the microenvironment of tumor initiation

Cytotechnology. 2023 Jun;75(3):243-253. doi: 10.1007/s10616-023-00575-1. Epub 2023 Apr 1.

Authors

Ghmkin Hassan^{1

2}, Said M Afify^{1

3}, Maram H Zahra^{1

4}, Hend M Nawara^{1

5}, Kazuki Kumon¹, Yoshiaki Iwasaki⁶, David S Salomon⁷, Akimasa Seno^{1

8

9}, Masaharu Seno^{1

9}

Affiliations

¹ Department of Cancer Stem Cell Engineering, Faculty of Interdisciplinary Science and Engineering in Health Systems, Institute of Academic and Research, Okayama University, Okayama, 700-8530 Japan.
² Department of Microbiology and Biochemistry, Faculty of Pharmacy, Damascus University, Damascus, Syria.
³ Division of Biochemistry, Chemistry Department, Faculty of Science, Menoufia University, Shebin El Koum, Menoufia 32511 Egypt.
⁴ Research Core for Interdisciplinary Sciences, Graduate School of Natural Science and Technology, Okayama University, Tsushima Naka, Kita, Okayama, 700-8530 Japan.
⁵ Department of Oncology, Lombardi Comprehensive Cancer Centre, Georgetown University, Washington, DC 20007 USA.
⁶ Health Service Center, Okayama University, Okayama, 700-8530 Japan.
⁷ Center for Cancer Research, National Cancer Institute, Frederick, MD 21702 USA.
⁸ The Laboratory of Natural Food and Medicine, Co, Ltd., Okayama, 700-8530 Japan.
⁹ R&D Center, Katayama Chemicals Ind., Co. Ltd, 4.1.7 Ina, Minoh, Osaka, 562-0015 Japan.

Abstract

Induced pluripotent stem cells (iPSCs) are useful tools for modeling diseases and developing personalized medicine. We have been developing cancer stem cells (CSCs) from iPSCs with conditioned medium (CM) of cancer-derived cells as the mimicry of the microenvironment of tumor initiation. However, the conversion of human iPSCs has not always been efficient with only CM. In this study, human iPSCs reprogrammed from monocytes of healthy volunteers were cultured in a media containing 50% of the CM from human pancreatic cancer derived BxPC3 cells supplemented with a MEK inhibitor (AZD6244) and a GSK-3α/β inhibitor (CHIR99021). The survived cells were assessed for the characteristics of CSCs in vitro and in vivo. As a result, they exhibited CSC phenotypes of self-renewal, differentiation, and malignant tumorigenicity. Primary culture of the malignant tumors of the converted cells exhibited the elevated expression of CSC related genes CD44, CD24 and EPCAM maintaining the expression of stemness genes. In conclusion, the inhibition of GSK-3α/β and MEK and the microenvironment of tumor initiation mimicked by the CM can convert human normal stem cells into CSCs. This study could provide insights into establishing potentially novel personalized cancer models which could help investigate the tumor initiation and screening of personalized therapies on CSCs.

Supplementary information: The online version contains supplementary material available at 10.1007/s10616-023-00575-1.

Keywords: Cancer stem cells; Human iPSCs; Signal pathway inhibitors; Tumor initiation.